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Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin

Decellularized membranes (DM) were obtained from bovine amniotic membranes (BAM) using four different decellularization protocols, based on physical, chemical, and mechanical treatment. The new material was used as a biological scaffold for in vitro skin cell culture. The DM were characterized using...

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Autores principales: Villamil Ballesteros, Andrea Catalina, Segura Puello, Hugo Ramiro, Lopez-Garcia, Jorge Andres, Bernal-Ballen, Andres, Nieto Mosquera, Diana Lorena, Muñoz Forero, Diana Milena, Segura Charry, Juan Sebastián, Neira Bejarano, Yuli Alexandra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7182835/
https://www.ncbi.nlm.nih.gov/pubmed/32151022
http://dx.doi.org/10.3390/polym12030590
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author Villamil Ballesteros, Andrea Catalina
Segura Puello, Hugo Ramiro
Lopez-Garcia, Jorge Andres
Bernal-Ballen, Andres
Nieto Mosquera, Diana Lorena
Muñoz Forero, Diana Milena
Segura Charry, Juan Sebastián
Neira Bejarano, Yuli Alexandra
author_facet Villamil Ballesteros, Andrea Catalina
Segura Puello, Hugo Ramiro
Lopez-Garcia, Jorge Andres
Bernal-Ballen, Andres
Nieto Mosquera, Diana Lorena
Muñoz Forero, Diana Milena
Segura Charry, Juan Sebastián
Neira Bejarano, Yuli Alexandra
author_sort Villamil Ballesteros, Andrea Catalina
collection PubMed
description Decellularized membranes (DM) were obtained from bovine amniotic membranes (BAM) using four different decellularization protocols, based on physical, chemical, and mechanical treatment. The new material was used as a biological scaffold for in vitro skin cell culture. The DM were characterized using hematoxylin-eosin assay, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR-ATR), and differential scanning calorimetry (DSC). The in vitro cytotoxicity of DM was evaluated using MTT. The efficacy of decellularization process was assessed through DNA quantification and electrophoresis. All the used protocols showed a high effectiveness in terms of elimination of native cells, confirmed by DNA extraction and quantification, electrophoresis, and SEM, although protocol IV removes the cellular contents and preserve the native extracellular matrix (ECM) architecture which it can be considered as the most effective in terms of decellularization. FTIR-ATR and DSC on the other hand, revealed the effects of decellularization on the biochemical composition of the matrices. There was no cytotoxicity and the biological matrices obtained were a source of collagen for recellularization. The matrices of protocols I, II, and III were degraded at day 21 of cell culture, forming a gel. The biocompatibility in vitro was demonstrated; hence these matrices may be deemed as potential scaffold for epithelial tissue regeneration.
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spelling pubmed-71828352020-05-01 Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin Villamil Ballesteros, Andrea Catalina Segura Puello, Hugo Ramiro Lopez-Garcia, Jorge Andres Bernal-Ballen, Andres Nieto Mosquera, Diana Lorena Muñoz Forero, Diana Milena Segura Charry, Juan Sebastián Neira Bejarano, Yuli Alexandra Polymers (Basel) Article Decellularized membranes (DM) were obtained from bovine amniotic membranes (BAM) using four different decellularization protocols, based on physical, chemical, and mechanical treatment. The new material was used as a biological scaffold for in vitro skin cell culture. The DM were characterized using hematoxylin-eosin assay, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR-ATR), and differential scanning calorimetry (DSC). The in vitro cytotoxicity of DM was evaluated using MTT. The efficacy of decellularization process was assessed through DNA quantification and electrophoresis. All the used protocols showed a high effectiveness in terms of elimination of native cells, confirmed by DNA extraction and quantification, electrophoresis, and SEM, although protocol IV removes the cellular contents and preserve the native extracellular matrix (ECM) architecture which it can be considered as the most effective in terms of decellularization. FTIR-ATR and DSC on the other hand, revealed the effects of decellularization on the biochemical composition of the matrices. There was no cytotoxicity and the biological matrices obtained were a source of collagen for recellularization. The matrices of protocols I, II, and III were degraded at day 21 of cell culture, forming a gel. The biocompatibility in vitro was demonstrated; hence these matrices may be deemed as potential scaffold for epithelial tissue regeneration. MDPI 2020-03-05 /pmc/articles/PMC7182835/ /pubmed/32151022 http://dx.doi.org/10.3390/polym12030590 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Villamil Ballesteros, Andrea Catalina
Segura Puello, Hugo Ramiro
Lopez-Garcia, Jorge Andres
Bernal-Ballen, Andres
Nieto Mosquera, Diana Lorena
Muñoz Forero, Diana Milena
Segura Charry, Juan Sebastián
Neira Bejarano, Yuli Alexandra
Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin
title Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin
title_full Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin
title_fullStr Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin
title_full_unstemmed Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin
title_short Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin
title_sort bovine decellularized amniotic membrane: extracellular matrix as scaffold for mammalian skin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7182835/
https://www.ncbi.nlm.nih.gov/pubmed/32151022
http://dx.doi.org/10.3390/polym12030590
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